新型生物催化剂的定向进化

诸平

在学术和工业实验室中越来越多地使用定向进化来修饰和改进重要的生物催化剂。定向进化改善了蛋白质和酶的功能，并在有机合成和生物技术中得到了应用。但是，存在多个缺点，使得研究人员无法充分利用这种方法的潜力。由于常见的基于PCR的诱变技术存在问题，因此迫切需要开发有效的筛选和选择工具来改进生物催化剂。科学家可以通过使用固相DNA合成技术来生成组合变体文库来减轻PCR的困难。这种方法减少了密码子偏倚，避免了序列错误，并鉴定了具有改进特性的新型突变体。

欲了解更多信息，可以从Twist Bioscience下载相关文献，以了解科学家如何在具有有效基因组装功能的硅芯片上使用高保真固相化学基因合成技术进​​行蛋白质发现；也可以注意浏览其它相关文献。

F. H. Arnold, A. A. Volkov. Directed evolution of biocatalysts. Curr Opin Chem Biol, 1999 Feb; 3(1): 54-59. DOI: 10.1016/s1367-5931(99)80010-6.

Abstract

Directed evolution is being used increasingly in academic and industrial laboratories to modify and improve important biocatalysts. Significant advances during this period of review include compartmentalization of genes and the in vitro translation apparatus in emulsions, as well as several impressive demonstrations of catalyst improvement. Shuffling of homologous genes offers a new way to utilize natural diversity in the evolution of novel catalysts.

Meng Wang, Tong Si, Huimin Zhao. Biocatalyst Development by Directed Evolution. Bioresour Technol. 2012 Jul; 115C: 117–125. Published online 2012 Jan 21. doi: 10.1016/j.biortech.2012.01.054

Abstract

Biocatalysis has emerged as a great addition to traditional chemical processes for production of bulk chemicals and pharmaceuticals. To overcome the limitations of naturally occurring enzymes, directed evolution has become the most important tool for improving critical traits of biocatalysts such as thermostability, activity, selectivity, and tolerance towards organic solvents for industrial applications. Recent advances in mutant library creation and high-throughput screening have greatly facilitated the engineering of novel and improved biocatalysts. This review provides an update of the recent developments in the use of directed evolution to engineer biocatalysts for practical applications.